Nanophotometer

The NanoPhotometer is a UV/Vis nano-volume spectrophotometer for quantitative and qualitative analysis of nucleic acid and protein samples in a submicroliter volume range. After the first and second generation (released 2006 and 2010) the third generation of the instrument, the NanoPhotometer P-Class, was released on October 24, 2011. The NanoPhotometer is used in many different scientific areas[1] like Zoology,[2] Plant Biology,[3] Microbiology,[4] Endocrinology,[5] Neuroscience[6] and Forensic.[7] Numerous state of the art techniques like Polymerase Chain Reaction (PCR),[8][9] Quantitative Real Time PCR (qPCR),[10] DNA Microarray Analysis[11][12][13] and Single-nucleotide Polymorphism (SNP)[14] require DNA, RNA and/or protein quantification of valuable samples. Only small volumes can be spared for this basic analysis. Therefore, a technique which enables UV/Vis quantification with minimal sample volumes has to be chosen. The NanoPhotometer requires a minimum volume of 0.3 µl. The wavelength range is between 190 – 1,100 nm. The detection range covers a broad region and is between 2 and 18,750 ng/μl for dsDNA and 0.08 and 543 mg/ml for protein (BSA). The NanoPhotometer offers small volume and cuvette capability.

Contents

Principle of low volume UV/Vis quantification

To ensure reliable quantitative and qualitative analysis of the sample an exactly defined pathway of the light through the sample is required. This is afforded by the patented Sample Compression Technology™ (patent numbers: DE 102004023178 and EP 1 910 807) of the NanoPhotometer. By choosing different lids, pathlengths between 0.04 mm and 2 mm are available. According to the Lambert Beer Law a reduction of the pathlength, compared to a standard 1 cm cuvette, results in a virtual dilution of the sample. In case of the NanoPhotometer dilution factors between 1/250 and 1/5 are available. Therefore, most measurements can be made with undiluted samples and valuable samples can be retrieved after the measurement. In addition the Sample Compression Technology™ avoids recalibrations of the instrument.

Low-volume quantification step by step

  1. For precise measurements accurate homogeneity of the sample is required. The integrated low vibration vortexer ensures homogeneous samples and consistent readings.
  2. The sample is pipetted onto the centre of the measuring window.
  3. Measuring chamber is covered by the appropriate lid.
  4. After measuring, sample can be retrieved with a pipette. The measuring window can be easily cleaned with a fluff-free tissue.

References

  1. ^ Scientific publications, http://quertle.info/search;jsessionid=CDB92762473EFB34D2830A90AD95B530?s=r&query=nanophotometer&so_a=&so_j=
  2. ^ Hiroyuki Kaiya et al. (2009), Identification and Genomic Sequence of a Ghrelin Receptor (GHS-R)-like Receptor in the Mozambique Tilapia, Oreochromis mossambicus. Zoological Science: Vol. 26, Issue 5, pg(s) 330-337, http://www.bioone.org/doi/abs/10.2108/zsj.26.330
  3. ^ Binod B. S. and Birendra P. S. (2009), Isolation, identification and expression analysis of salt-induced genes in Suaeda maritima, a natural halophyte, using PCR-based suppression subtractive hybridization. BMC Plant Biology, Vol. 9, Issue 69, pg(s) 1-25, http://www.biomedcentral.com/content/pdf/1471-2229-9-69.pdf
  4. ^ Kuo-Hsiang Tang et al (2009), Role of the AcsF Protein in Chloroflexus aurantiacus. J. Bacteriol., Vol.191, Issue 11, pg(s) 3580-3587, http://jb.asm.org/content/191/11/3580.full.pdf+html
  5. ^ Sarath Chandra Peddu et al. (2009), Pre- and postprandial effects on ghrelin signaling in the brain and on the GH/IGF-I axis in the Mozambique tilapia (Oreochromis mossambicus). General and Comparative Endocrinology, Volume 161, Issue 3, pg(s) 412-418, http://www.sciencedirect.com/science/article/pii/S0016648009000719
  6. ^ Naomi L. Cook et al. (2009), Validation of reference genes for normalization of real-time quantitative RT-PCR data in traumatic brain injury. Journal of Neuroscience Research, Vol 87, Issue 1, pg(s) 34 – 41, http://onlinelibrary.wiley.com/doi/10.1002/jnr.21846/abstract
  7. ^ Erin K. Hanson and Jack Ballantyne (2010), A Blue Spectral Shift of the Hemoglobin Soret Band Correlates with the Age (Time Since Deposition) of Dried Bloodstains, PLoS One, 5(9): e12830, http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2942901/
  8. ^ Xiaokang Wu et al. (2010) Molecular Characterisation of the Faecal Microbiota in Patients with Type II Diabetes, Current Microbiology, Volume 61, Number 1, http://www.springerlink.com/content/30340mg048m54651/
  9. ^ Matthias Brock et al. (2008) Bioluminescent Aspergillus fumigatus, a New Tool for Drug Efficiency Testing and In Vivo Monitoring of Invasive Aspergillosis. Appl Environ Microbiol. 2008 November: Vol 74, Issue 22, pg(s) 7023–7035 http://aem.asm.org/content/74/22/7023.full.pdf+html
  10. ^ Naomi L. Cook et al. (2009), Validation of reference genes for normalization of real-time quantitative RT-PCR data in traumatic brain injury. Journal of Neuroscience Research, Vol 87, Issue 1, pg(s) 34 – 41, http://onlinelibrary.wiley.com/doi/10.1002/jnr.21846/abstract
  11. ^ Anuj Kumar Gupta et al. (2010), Microarray Analysis of Efflux Pump Genes in Multidrug-Resistant Mycobacterium tuberculosis During Stress Induced by Common Anti-Tuberculous Drugs, Microbial Drug Resistance, Volume 16, Issue1, http://www.liebertonline.com/doi/abs/10.1089/mdr.2009.0054
  12. ^ David Velázquez-Fernández (2005), Differential RNA Expression in Benign and Malignant Adrenocortical Tumours, Dissertation, Karolinska University Press, Stockholm Sweden, ISBN 91-7140-578-X, http://diss.kib.ki.se/2005/91-7140-578-X/thesis.pdf
  13. ^ Jana Flügge (2008), Haplotypen des Vitamin-D-Rezeptor-Gens als Suszeptibilitätsfaktoren für das kolorektale Karzinom sowie In-vitro-Expressionsanalyse Vitamin-D-regulierter Gene, Dissertation Fachbereich Biologie, Chemie, Pharmazie der Freien Universität Berlin, http://www.diss.fu-berlin.de/diss/servlets/MCRFileNodeServlet/FUDISS_derivate_000000005286/Dissertation-JanaFl%C3%BCgge.pdf?hosts
  14. ^ Ludwig Czibere (2008), Assessing the complex nature of behavior: Sequence-based and transcriptomic analyses in a mouse model of extremes in trait anxiety, Dissertation, Faculty of Biology, Ludwig-Maximilians-University Munich, http://edoc.ub.uni-muenchen.de/10207/1/Czibere_Ludwig.pdf

External links

Official NanoPhotometer Web Site

NanoPhotometer Brochure

NanoPhotometer Manual

NanoPhotometer Testimonials